c-kit (CD117) Antibody Staining Protocol for Immunohistochemistry

c-kit Oncoprotein (CD117) Antibody Staining Protocol for Immunohistochemistry

Description: The c-kit proto-oncogene encodes a transmembrane tyrosine kinase receptor, c-kit (CD117) which is closely related to the platelet derived growth factor family. c-kit plays a role during haematopoiesis, gametogenesis and melanogenesis.

Primary Antibody

Name: c-kit Oncoprotein (CD117) Antibody

Clone: 57A5D8, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-cKIT

Dilution: 1:200 - 1:400 using IHC-Tek^TM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 minutes/room temperature

Antigen Retrieval

Device: No antigen retrieval is needed

Buffer/pH value: N/A

Heat/Cool Temperature: N/A

Heat/Cool Time: N/A

Detection Methods

Standard Method: ABC Method or LSAB Method

Enhanced Method: Polymeric Methods

Chromogen Substrate

Reagent: DAB

Incubation Time/Temperature: 1-3 minutes/room temperature

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

Staining Pattern: Membrane/cytoplasmic

Images: Search image

Additional Information:

Tissue Type: Skin

Fixation: Acetone fixed frozen sections.

Positive Control: Normal skin, epidermal melanocytes.

Negative Control: Omit primary antibody, isotype control or absorption control

Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary

Notes:

Not tested on paraffin sections

References: